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来自羊种布鲁氏菌的重组Omp31单独或与粗糙脂多糖联合使用时,可诱导BALB/c小鼠对绵羊布鲁氏菌感染产生保护作用。

The recombinant Omp31 from Brucella melitensis alone or associated with rough lipopolysaccharide induces protection against Brucella ovis infection in BALB/c mice.

作者信息

Estein Silvia M, Cassataro Juliana, Vizcaíno Nieves, Zygmunt Michel S, Cloeckaert Axel, Bowden Raúl A

机构信息

Laboratorio de Inmunoquímica y Biotecnología, Departamento de Sanidad Animal y Medicina Preventiva, Facultad de Ciencias Veterinarias, UNICEN, 7000 Tandil, Argentina.

出版信息

Microbes Infect. 2003 Feb;5(2):85-93. doi: 10.1016/s1286-4579(02)00075-8.

Abstract

Immunogenicity and protective activity against Brucella ovis of detergent-extracted recombinant Omp31 (rOmp31 extract) from Brucella melitensis produced in Escherichia coli, purified rough lipopolysaccharide from B. ovis (R-LPS) and a mixture of rOmp31 extract and R-LPS (rOmp31 extract + R-LPS) were assessed in BALB/c mice. The experimental vaccines were compared with a hot saline extract (HS extract) from B. ovis mainly composed of outer membrane proteins (OMPs) and R-LPS, and known to be protective in mice against a B. ovis infection. Serum antibodies to Omp31 and R-LPS were detected in the corresponding mice using Western blotting with B. ovis whole-cell lysates and ELISA with purified antigens. Protection was evaluated by comparing the levels of infection in the spleens of vaccinated mice challenged with B. ovis. A significantly lower number of B. ovis colony-forming units in spleens relative to unimmunized (saline injected) controls were considered as protection. Mice immunized with rOmp31 extract or rOmp31 extract mixed with R-LPS developed antibodies that bound to the B. ovis surface with similar titers. Vaccination with rOmp31 extract plus R-LPS provided the best protection level, which was comparable with that given by HS extract. Similar protection was also obtained with rOmp31 extract alone and, to a lesser degree, with R-LPS. Comparisons between groups showed that an extract from E. coli-pUC19 (devoid of Omp31) provided no protection relative to either HS extract, rOmp31 extract or rOmp31 extract mixed with R-LPS. In conclusion, the recombinant Omp31 associated or not with B. ovis R-LPS, could be an interesting candidate for a subcellular vaccine against B. ovis infection.

摘要

在BALB/c小鼠中评估了用大肠杆菌生产的来自布鲁氏菌的去污剂提取重组Omp31(rOmp31提取物)、来自绵羊布鲁氏菌的纯化粗糙脂多糖(R-LPS)以及rOmp31提取物与R-LPS的混合物(rOmp31提取物+R-LPS)对绵羊布鲁氏菌的免疫原性和保护活性。将实验疫苗与主要由外膜蛋白(OMPs)和R-LPS组成的绵羊布鲁氏菌热盐水提取物(HS提取物)进行比较,已知该提取物对小鼠抵抗绵羊布鲁氏菌感染具有保护作用。使用绵羊布鲁氏菌全细胞裂解物进行蛋白质印迹法以及使用纯化抗原进行酶联免疫吸附测定法,检测相应小鼠中针对Omp31和R-LPS的血清抗体。通过比较用绵羊布鲁氏菌攻击的接种疫苗小鼠脾脏中的感染水平来评估保护作用。相对于未免疫(注射生理盐水)对照,脾脏中绵羊布鲁氏菌菌落形成单位数量显著降低被视为具有保护作用。用rOmp31提取物或与R-LPS混合的rOmp31提取物免疫的小鼠产生了以相似滴度与绵羊布鲁氏菌表面结合的抗体。用rOmp31提取物加R-LPS进行疫苗接种提供了最佳保护水平,与HS提取物提供的保护水平相当。单独使用rOmp31提取物以及程度较小地使用R-LPS也获得了类似的保护作用。组间比较表明,来自大肠杆菌-pUC19(不含Omp31)的提取物相对于HS提取物、rOmp31提取物或与R-LPS混合的rOmp31提取物均无保护作用。总之,与绵羊布鲁氏菌R-LPS相关或不相关的重组Omp31,可能是预防绵羊布鲁氏菌感染的亚细胞疫苗的一个有吸引力的候选物。

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